Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.

Background Adeno-associated virus-based vector is one of most efficient vehicles.It presents with a long-term and efficient transfer and expression of therapeutic genes with minimal toxicity.But its delayed-and low-efficient transgene expression limits the application of AAV vector.To explore an improving method of AAV infecting RPE cells is the hot spot. Objeetive Present study was to investigate whether adeno-associated virus (AAV)combined with low dose non-replieable adenovirus(Ad-null)can enhance its infection efficieney on RPE ceils in vitro. Methods Human RPE cells were isolated from the donate eyeballs under the approval of the Ethic Committee of this hospital.The cells were cultured in DMEM containing 10%fetal bovine serum.AAV particles with enhanced green fluorescence protein(EGFP)were added into the medium alone or in combination with different amount of adenovirus for 30 days.The cells were detected under the fluorescence microscope,and the protein expression levels of report gene EGFP in RPE cells were analyzed with Western blotting assay. Results Melanin granules could be found in cultured RPE cells.EGFP was expressed in RPE cells at 2 days after AAV-EGFP infection and peaked at 12 days and remained for about 3-week duration,showing the green influorescence under the influorescence mwroscope.After the cells were infected by AAV2-EGFP with 0.01 to 1000 MOI Ad-null respectively,the number of cells with green influorescence was obviously increased with the enhanced infiuorescence intensity.The enhance of the infection efficiency began in the 0.1 MOI Ad-null group and peaked in 10 MOI Ad-null group.Dead cells were exhibited in the 100 or more MOI Ad-nulor group.Western blotting assay demonstrated that the protein expression level of EGFP in RPE cells enhanced significantly in 1 and 10 MOI Ad-null groups compared with only AAV infection group. Conclusion These finding suggested that the infection efficiency of AAV can be improved significantly when it is used with low dose Ad-null in vitro.This offers a basis for further study of gene therapy.

Eye Enucleation ; Humans ; Infant ; Retinal Neoplasms ; surgery ; Retinoblastoma ; surgery ; Vitrectomy

Retinoblastoma (RB) is one of the most common primary intraocular malignant tumors of children. With improvement of the medical care, the goal of RB treatment is saving the eyeball and even the vision. In the process of globe-salvaging treatment, complicated cataract has been a serious problem which affects the treatment and follow-up. It is important to operate a cataract surgery timely and safely. In this paper related research advances were summarized.

Intra-arterial chemotherapy (IAC) refers to the injection of chemotherapeutics into patients' eyes through arteria ophthalmica in order to local chemotherapy for retinoblastoma (RB).The common drugs in the treatment of RB include melphalan,topotecan and carboplatin.IAC can form the high-concentration drug effects on local areas.It can effectively kill tumors and significantly reduce the occurrence rate of complications after intravenous chemotherapy.Application of IAC can cure patients with RB in B,C,D and E stage in international criteria,which can effectively improve eye protection rate.Complications of IAC include eyelids edema,chemosis,blepharoptosis,ocular motility disorders,vitreous hemorrhage,ischemia of choroid or retina,transient myelosuppression,etc.It should be further conduction of long-term and large-sample cases studies on how to improve curative effects and reduce serious eye complications.

OBJECTIVETo prospectively analyze the correlation of MRI image characteristics with histopathological findings in pancreatic head carcinoma in vivo and ex vivo.

METHODSSixteen patients with pancreatic head carcinoma were examined by 1.5T MRI. MR sequences included pre-and post-contrast dynamic 2D FLASH T1WI, TSE T2WI + fat suppressed (FS). All fresh specimens were scanned again within an hour after resection during the Whipple procedure, and were then cut into slices along the direction of MRI scan. The MRI image characteristics were compared with gross and histopathologic findings of the specimens.

RESULTSThe tumor size ranged from 1.5 cm x 2.0 cm to 4.8 cm x 4.2 cm with a mean value of 3.6 cm x 3.1 cm. The MRI findings in vivo showed hypointensity in 14 lesions and isointensity in 2, and on 2D FLASH T1WI and TSE T2WI, all lesions displayed mixed intensity except 3 lesions with iso-intensity. During post-contrast dynamic pancreatic parenchymal phase, 15 lesions showed mild enhancement and 11 lesions had ring enhancement sign. Nine lesions displayed progressive irregular moderate enhancement during post-contrast dynamic hepatic phase. Four lesions showed enhancement with iso- and hyperintensity in post-contrast dynamic delayed phase. The MRI findings demonstrated that all lesions ex vivo had hypointensity on 2D FLASH T1 WI and mixed intensity on TSE T2WI + FS. Tumor tissues mainly displayed hypointensity, and the area containing tumor and inflammatory tissue showed iso-intensity on 2D FLASH T1WI. On T2WI + FS, the fibrosis proportion displayed hypo or isointensity, while the tumor and chronic inflammatory tissue demonstrated slight hyperintensity, and the zones of mucous degeneration or pancreatic ductal dilatation displayed hyper-intensity. The ring enhancement of tumor was caused by multiple factors and no obvious enhancement within tumor and fibrosis area were observed during post-contrast dynamic pancreatic parenchymal phase. Progressive irregular enhancement originated from desmoplastic reaction within the tumor during post-contrast dynamic hepatic phase and delayed phase, respectively.

CONCLUSIONPancreatic head carcinomas contain various kinds of tissues. MRI can reveal these pathologic characteristics. No obvious hemorrhage and necrosis within the tumor were observed in this series.

Adult ; Aged ; Female ; Humans ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Pancreas ; pathology ; Pancreatectomy ; Pancreatic Neoplasms ; diagnosis ; pathology ; surgery ; Prospective Studies

OBJECTIVETo investigate the CT and MRI manifestatitions of intraductal papillary mucinous neoplasm (IPMN) of the pancreas.

METHODSBoth clinical and imaging data of 12 pathologically confirmed intraductal papillary mucinous neoplasm, of the pancreas were retrospectively analyzed.

RESULTSThe pancreatic IPMN can be classified into two types based on CT image: the branch duct IPMN (n=7) originated from the head and uncinate process of the pancreas. The tumor consisted of lobulated or clustered small cyst lesions with septa among them, the wall and septa can be enhanced; the combined IPMN (n=5) involved branch ducts of the uncinate process as well as the main pancreatic ducts with dilatation (diameter: 4-7 mm), one of these involved the branch ducts along the pancreatic body. The pancreatic IPMN was mainly found in elderly patient with a chief clinical symptoms of abdominal pain and/or pancreatitis.

CONCLUSIONThe intraductal papillary mucinous neoplasm of the pancreas enjoys specific features in CT and MRI image, which are helpful to the diagnosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Pancreatic Ductal ; diagnosis ; diagnostic imaging ; Carcinoma, Papillary ; diagnosis ; diagnostic imaging ; Cystadenocarcinoma, Mucinous ; diagnosis ; diagnostic imaging ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pancreatic Ducts ; diagnostic imaging ; pathology ; Pancreatic Neoplasms ; diagnosis ; diagnostic imaging ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, X-Ray Computed

In order to clarify the chemical constituents in Qiliqiangxin capsule, a rapid ultra-performance liquid chromatography/orthogonal acceleration time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established. Forty peaks were identified on line using this method. The herbal sources of these peaks were assigned. The results implied that triterpenoid saponins, flavonoid glycosides, C21-steroids and phenolic acids were included in the main components of Qiliqiangxin capsule. The method is simple and rapid for elucidation of the constituents of Qiliqiangxin capsule and the results are useful for the quality control of Qiliqiangxin capsule.
Capsules ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Flavones ; analysis ; Ginsenosides ; analysis ; Glycosides ; analysis ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Saponins ; analysis ; Spectrometry, Mass, Electrospray Ionization ; Steroids ; analysis ; Tandem Mass Spectrometry ; Triterpenes ; analysis

OBJECTIVETo explore the methods of diagnosis and surgical treatment for patients with back pain originating from endplate.

METHODSAll patients received examinations of radiography, CT, and MR imaging. Pain level of disc was decided by discography in each patient. The principal outcome judgment were pain and disability, and the efficacy of surgical treatment was assessed by visual analog scale (VAS) for pain, and the Oswestry disability index (ODI) for functional recovery.

RESULTSAll patients with a diagnosis of back pain originating from endplate according to discography were treated with anterior or posterior fusion surgery. The mean follow-up period was three years and five months (from 2 to 6 years). Among of the 21 patients, 20 (95%) reported a disappearance or marked alleviation of low back pain, and experienced a definite improvement in physical function. A statistically significant and clinically meaningful improvement on the ODI and the VAS scores were obtained in the patients with chronic low back pain originating from endplate (P = 0.0001) after treatment.

CONCLUSIONSThe study suggests that the discography and fusion surgery may be very effective methods for the diagnosis and treatment of chronic back pain originating from endplate respectively.

Adult ; Female ; Follow-Up Studies ; Humans ; Low Back Pain ; diagnosis ; etiology ; surgery ; Male ; Middle Aged ; Spinal Fusion ; methods ; Spinal Injuries ; complications ; Treatment Outcome

OBJECTIVETo investigate the role of MRI in evaluating the peripancreatic vessel invasion and resectability of pancreatic carcinoma based on the comparison of MRI image with surgical exploration, and try to establish the criteria for assessment of the sensitivity, specificity and accuracy of resectability.

METHODSForty-one pancreatic carcinoma patients confirmed by pathology received preoperative plain and contrast enhanced MRI scan, and 37 of them had additional coronal MRA scan. Peripancreatic vessel invasion was preoperatively assessed based on MRI features, and the vessel invasion degree from the uninvolved to the severely involved was divided into 6 grades represented by 1, 2a, 2b, 3a, 3b and 4, respectively. Compared with the findings during the surgery, the sensitivity and specificity of each vessel invasion grade were studied and the receiver operator characteristic curve (ROC) was drawn. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of resectability evaluation based on 2 and 2a degree were calculated respectively. The resectability of involved arteries and veins of grade 2 were also analyzed.

RESULTSOf the 41 patients, 22 had resectable tumor, with 20 curative resection and 2 palliative. Compared with the findings during surgery, seven vessels including three arteries and four veins were not correctly interpreted by MRI. If grade 1,2a,2b,3a and 3b was used as the resectable standard,respectively, the sensitivity to predict the unresectbility was 78.3%, 84.8%, 67.4%, 56.5% and 47.8%, respectively. Receiver operator characteristic curve demonstrated that grade 2a was the optimal critical point. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of grade 2a in predicting the unresectbility were 84.8%, 98.5%, 92.9%, 96.6% and 95.9%.

CONCLUSIONOur data showed that grade 2a (tumor involvement < 2 cm long and < 1/2 circumference of the vessel) may be more sensitive and accurate in predicting the resectability, which may be considered as the line of demarcation between the respectable and unresectable cases in clinical practice.

Adult ; Aged ; Carcinoma, Pancreatic Ductal ; diagnosis ; pathology ; surgery ; Celiac Artery ; pathology ; Cholangiopancreatography, Magnetic Resonance ; Female ; Hepatic Artery ; pathology ; Humans ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Male ; Mesenteric Artery, Superior ; pathology ; Mesenteric Veins ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Pancreas ; blood supply ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; diagnosis ; pathology ; surgery ; Portal Vein ; pathology ; Predictive Value of Tests ; Sensitivity and Specificity